In 'Vitro' Whole Embryo Culture: Methods Development

摘要

Whole embryo culture of Mammalian embryos (WEC) has been used for a number of years as an approach to basic science studies but only recently have investigators begun to examine its potential as a screen of teratogenesis. A laboratory was established utilizing the basic techniques for WEC that have been developed over the past 15 years. This included aspects of serum collection and processing, embryo explantation and culture, and morphological evaluation. The technique for embryo explantation was developed to provide embryos in which the visceral yolk sac and ectoplacental cone remained intact. Day 10 embryos, cultured over a 48-hour period, were evaluated for both viability and development using a morphological scoring system. The gassing regimen most compatible with a high percent viability and appropriate morphological development was one in which the initial concentration of oxygen was 5%, increasing to 95% during the second day of incubation. This led to a viability of 92% over the 48-hour culture period and a morphological score appropriate to a day 12 embryo in vivo. This technique will be applied in future experiments designed to evaluate developmental parameters in WEC and their use in validating the system as a potential screen for teratogens.